CN219652999U - Microorganism triangular flask culture apparatus - Google Patents
Microorganism triangular flask culture apparatus Download PDFInfo
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- CN219652999U CN219652999U CN202223534733.2U CN202223534733U CN219652999U CN 219652999 U CN219652999 U CN 219652999U CN 202223534733 U CN202223534733 U CN 202223534733U CN 219652999 U CN219652999 U CN 219652999U
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- pipeline
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- flask
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- 244000005700 microbiome Species 0.000 title claims abstract description 15
- 239000011521 glass Substances 0.000 claims abstract description 69
- 238000000855 fermentation Methods 0.000 claims abstract description 22
- 230000004151 fermentation Effects 0.000 claims abstract description 22
- 230000000813 microbial effect Effects 0.000 claims abstract description 19
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000000741 silica gel Substances 0.000 claims abstract description 18
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 18
- 239000007787 solid Substances 0.000 claims abstract description 15
- 239000012528 membrane Substances 0.000 claims description 20
- 230000001954 sterilising effect Effects 0.000 claims description 15
- 229910001220 stainless steel Inorganic materials 0.000 claims description 9
- 239000010935 stainless steel Substances 0.000 claims description 9
- 230000000844 anti-bacterial effect Effects 0.000 claims description 7
- 238000007789 sealing Methods 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 241000894006 Bacteria Species 0.000 abstract description 26
- 230000002572 peristaltic effect Effects 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 6
- 239000013589 supplement Substances 0.000 abstract description 3
- 230000001502 supplementing effect Effects 0.000 abstract description 3
- 238000000034 method Methods 0.000 description 15
- 238000011081 inoculation Methods 0.000 description 8
- 238000005070 sampling Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 240000006024 Lactobacillus plantarum Species 0.000 description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 4
- 239000000306 component Substances 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 241000233866 Fungi Species 0.000 description 3
- 238000011109 contamination Methods 0.000 description 3
- 241000193749 Bacillus coagulans Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000019764 Soybean Meal Nutrition 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 229940054340 bacillus coagulans Drugs 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 150000002303 glucose derivatives Chemical class 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 239000004455 soybean meal Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000012840 feeding operation Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229940081969 saccharomyces cerevisiae Drugs 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model provides a microorganism triangular flask culture device which comprises a triangular flask, wherein the triangular flask comprises a glass flask body, a glass pipeline and a solid glass support for supporting the glass pipeline, an opening is formed in one side wall of the flask body, one end of the glass pipeline extends to the bottom of the inside of the flask body through the opening, the other end of the glass pipeline is positioned outside the flask body, and a silica gel transparent hose or a feeding pipeline is sleeved on the glass pipeline. The microbial triangular flask culture device disclosed by the utility model can supplement materials and measure pH, is convenient to operate, is not easy to dye bacteria, has the advantages of the triangular flask and the fermentation tank, can be inoculated into the fermentation tank in a peristaltic pump feeding mode after the culture is finished, is not easy to dye bacteria, has a simple structure, is low in cost, is convenient to operate and carry, does not need to be matched with a material supplementing bottle and the like, and can be used for completing the regulation and control of microorganisms only by using a syringe and a connecting accessory.
Description
Technical Field
The utility model relates to the technical field of microorganism culture, in particular to a microorganism triangular flask culture device.
Background
The microbial triangular flask culture is a key step of microbial fermentation culture, and can provide reliable basis for optimizing culture medium components and regulating fermentation parameters in microbial industrial production. However, the existing triangular flask level microbial culture adopts an intermittent culture mode, namely the culture process does not need the regulation and control operations such as material supplementing, pH adjustment and the like, so that the characteristics of low bacterial concentration, weak bacterial activity and the like of the culture are caused; in addition, after the culture of the triangular flask is finished, the fermentation tank is inoculated by adopting a fire ring method, and the operation step has high risk of bacteria contamination; secondly, the existing triangular flask improvement mode has the characteristics that the existing triangular flask improvement mode is close to a fermentation tank and can be used for fed-batch culture, but the refitting device is complex, the operation and the culture are not convenient enough, and the portable multifunctional triangular flask has no characteristics.
In view of the above, the present utility model provides a microorganism flask culture device to solve the above technical problems.
Disclosure of Invention
Aiming at the defects of the prior art, the utility model provides the microbial triangular flask culture device which can supplement materials and measure pH, is convenient to operate, is not easy to dye bacteria, has the advantages of the triangular flask and the fermentation tank, can be inoculated into the fermentation tank in a peristaltic pump feeding mode after the culture is finished, and is not easy to dye bacteria.
The utility model adopts the following technical scheme for solving the technical problems:
the utility model provides a microorganism triangular flask culture apparatus, including the triangular flask, the triangular flask includes glass body, glass pipeline and is used for supporting solid glass support of glass pipeline, an opening has been seted up on the bottle wall of one side of glass body, one end of glass pipeline passes through the opening stretches to the inside bottom of glass body, and the other end is located the outside of glass body, glass pipeline cover is equipped with silica gel transparent hose or stream and adds the pipeline.
Further, the triangular flask is composed of a glass flask body with a conical bottom and an arc-shaped flask opening at the upper part, and a breathable antibacterial sealing film is sleeved on the flask opening.
Further, a degerming filter membrane, or a degerming filter membrane and a syringe, or a syringe is arranged on the silica gel transparent hose, and the degerming filter membrane is detachably connected with the syringe.
Further, the pore diameter of the degerming filter membrane is 0.2-0.25um.
Further, the feeding pipeline comprises a silica gel hose and a stainless steel needle, the stainless steel needle is inserted into a feeding interface of the fermentation tank, and the silica gel hose is sleeved on the glass pipeline.
Further, the solid glass support and the bottom of the glass bottle body are arranged in parallel.
Further, one end of the solid glass support is fixedly connected with the glass pipeline, and the other end of the solid glass support is fixedly connected with the inner bottle wall of the glass bottle body.
Further, the glass bottle body, the glass pipeline and the solid glass bracket are integrally formed.
Compared with the prior art, the utility model has the following technical advantages:
1. the microbial triangular flask culture device disclosed by the utility model can supplement materials and measure pH, is convenient to operate, is not easy to dye bacteria, has the advantages of the triangular flask and the fermentation tank, can be inoculated into the fermentation tank in a peristaltic pump feeding mode after the culture is finished, is not easy to dye bacteria, has a simple structure, is low in cost, is convenient to operate and carry, does not need to be matched with a material supplementing bottle and the like, and can be used for completing the regulation and control of microorganisms only by using a syringe and a connecting accessory.
2. The microbial triangular flask culture device provided by the utility model realizes the microbial culture process, detects and regulates the growth state, is not easy to stain bacteria, can improve the microbial growth performance of the triangular flask level, namely, increase the concentration of bacteria, improve the activity of bacteria or improve the content of products, and can be used for culturing various types of microorganisms such as saccharomycetes, lactobacillus and bacteria.
3. The microbial triangular flask culture device reduces the risk of bacteria contamination in the process of inoculating the triangular flask to the fermentation tank, can transfer more fragile strains, and is suitable for inoculation in a worse environment.
Drawings
FIG. 1 is a schematic view of the structure of a triangular flask according to the present utility model;
FIG. 2 is a schematic diagram of a flask-fitted feed line in accordance with the present utility model;
FIG. 3 is a schematic view of the structure of the triangular flask matched with the silica gel transparent hose in the utility model;
FIG. 4 is a cross-sectional view of the triangular flask in combination with a sterilizing filter and syringe according to the present utility model;
FIG. 5 is a schematic view of the structure of the triangular flask matched with the silica gel transparent hose and the syringe in the utility model;
FIG. 6 is a schematic structural view of an antibacterial sealing film according to the utility model;
FIG. 7 is a schematic view of the structure of the sterilizing filter according to the present utility model;
wherein the reference numerals are as follows:
a glass bottle body 1; a glass tube 2; a solid glass support 3; a bottle mouth 4; an antibacterial sealing film 5; a feeding pipeline 6; a silicone hose 61; stainless steel needle 62; a silicone transparent hose 7; a syringe 8; and a sterilizing filter membrane 9.
Detailed Description
The following description of the embodiments of the present utility model will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present utility model, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
In the description of the present utility model, it should be noted that the azimuth or positional relationship indicated by the terms "vertical", "upper", "lower", "horizontal", etc. are based on the azimuth or positional relationship shown in the drawings, and are merely for convenience of describing the present utility model and simplifying the description, and do not indicate or imply that the apparatus or element referred to must have a specific azimuth, be constructed and operated in a specific azimuth, and thus should not be construed as limiting the present utility model.
In the description of the present utility model, it should also be noted that, unless explicitly specified and limited otherwise, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present utility model can be understood by those of ordinary skill in the art according to the specific circumstances.
Example 1
Referring to fig. 1-7, this embodiment provides a microorganism triangular flask culture apparatus, including the triangular flask, the triangular flask includes glass body 1, glass pipeline 2 and is used for supporting the solid glass support 3 of glass pipeline 2, an opening has been seted up on the bottle wall of one side of glass body 1, one end of glass pipeline 2 passes through the opening stretches to the inside bottom of glass body 1, and the other end is located the outside of glass body 1, glass pipeline 2 cover is equipped with silica gel transparent hose 7 or stream and adds pipeline 6.
In the embodiment, the triangular flask is composed of a glass flask body 1 with a conical bottom and an arc-shaped flask mouth 4 at the upper part, and a breathable antibacterial sealing film 5 is sleeved on the flask mouth 4.
In this embodiment, the silica gel transparent hose 7 is provided with a sterilizing filter membrane 9, or the sterilizing filter membrane 9 and the syringe 8, or the syringe 8, and the sterilizing filter membrane 9 is detachably connected with the syringe 8.
In this embodiment, the pore size of the sterilizing filter membrane 9 is 0.2-0.25um, preferably, the pore size of the sterilizing filter membrane 8 is 0.22umm.
In this embodiment, the feeding pipeline 6 includes a silica gel hose 61 and a stainless steel needle 62, the stainless steel needle 62 is inserted into a feeding port of the fermentation tank, and the silica gel hose 61 is sleeved on the glass pipeline 2.
In this embodiment, the solid glass support 3 and the bottom of the glass bottle body 1 are disposed in parallel, one end of the solid glass support 3 is fixedly connected with the glass tube 2, and the other end is fixedly connected with the inner wall of the glass bottle body 1.
In this embodiment, the glass bottle body 1, the glass tube 2 and the solid glass bracket 3 are integrally formed.
The normal culture of microorganism is shown in figure 3, the antibacterial sealing film 5 is sleeved on the bottle mouth 4 of the triangular flask, the silica gel transparent hose 7 is sleeved on the glass pipeline 2 of the triangular flask, the thin end of the antibacterial filtering film 9 is sleeved on the injector 8, and the culture process is similar to that of a common triangular flask.
The working process of the microbial triangular flask culture device of the embodiment is as follows:
when pH detection and feeding operations are required, a graduated syringe 8 may be used to sample or feed through the sterile filter 8. For example, the pH may be monitored by sampling via the syringe 8, or the bacteria concentration may be monitored, and the bacteria concentration may be monitored, wherein the bacteria filter 9 is not removed if the feed components are soluble substances, as shown in fig. 4, but the bacteria filter 9 needs to be replaced after the feed is completed. If the feed ingredients have insoluble substances or the bacteria concentration is required to be measured in sampling, the filter membrane is removed as shown in figure 5, and the filter membrane is sleeved after the operation.
When the seed culture is finished, if the seed liquid is required to be transferred into the fermentation tank, the transparent silica gel hose 7 and the silica gel hose 61 of the feeding pipeline 6 can be connected as shown in fig. 2, then the stainless steel needle 62 of the feeding pipeline 6 is inserted into the feeding port of the fermentation tank, and then the seed liquid is pumped into the fermentation tank through a peristaltic pump.
It should be noted that the above operations all require aseptic operations, i.e. all media, bottles, and fittings used require a treatment of 20mmin at 121℃and the sampling and feeding processes are all performed in a sterile super clean bench. Inoculating from the triangular flask to the fermentation tank, connecting the feeding pipeline 6 and the bottle in an ultra-clean workbench, binding the stainless steel needle 62 of the feeding pipeline 6, and opening beside a fire ring and then inserting into the fermentation tank when transferring to the fermentation tank.
Example 2
In this example, the culture control and transfer inoculation operation of lactobacillus plantarum were performed using the microorganism flask culture apparatus in example 1 described above.
Selecting one strain of Lactobacillus plantarum, performing MRS culture medium liquid culture, and concentrating to 1×10 8 Fu/mL, 37 ℃, culture 24 hours. The common triangular flask culture process does not need any operation, the modified triangular flask is used for sampling and measuring pH respectively in culture for 6h, 12h and 18h, the pH is adjusted to 5.5-6.0 by adding 5mol/L sodium hydroxide into a syringe 8, and 1% sterile saturated glucose solution is supplemented. Since MRS medium, glucose and alkali solution are all soluble components, the matching mode of FIG. 4 can be selected in the sampling and feeding process, namely the sterilizing filter membrane 9 is not required to be removed, but the sterilizing filter membrane 9 needs to be replaced after the operation is finished.
After the culture is finished, the bacterial concentration of the common triangular flask is measured to be 2.0x10 9 The fungus concentration of the feeding triangular flask is 5.0X109 fu/mL, and the fungus concentration is about 2.5 times of that of the common triangular flask.
After the cultivation is finished, the cultivated seed liquid is inoculated to a fermentation tank respectively in an environment with more mixed bacteria, the seeds cultivated in a common triangular flask are inoculated by a fire ring method, mixed bacteria in the air can be introduced, the seeds cultivated in the improved triangular flask are inoculated by a peristaltic pump in a fed-batch mode, and the condition that the seeds are not infected by bacteria is verified to be fed-batch by the peristaltic pump in a fed-batch mode for many times, so that the effective rate is 100%.
Example 3
In this example, the culture control and transfer inoculation operation of lactobacillus plantarum were performed using the microorganism flask culture apparatus in example 1 described above.
Selecting a strain of Bacillus coagulans, culturing with 1% glucose, 2% soybean meal, 1% yeast extract, and 1% sodium chloride as culture medium, and concentrating to 2×10 8 u/mL, 37 ℃, 200r/min culture 24 hours.
The common triangular flask culture process does not need any operation, the modified triangular flask is used for sampling and measuring pH respectively in culture for 6h, 12h and 18h, 5mol/L sodium hydroxide or 5mol/L hydrochloric acid is added into the modified triangular flask by a syringe 8 to adjust the pH to 6.5-7.0, and 1% sterile saturated glucose solution is supplemented. The method matched with FIG. 5 can be selected in the sampling and feeding processes because insoluble soybean meal exists in the culture medium components, namely the sterilizing filter membrane 9 is removed, but the sterilizing filter membrane 9 is sleeved after the operation is finished.
After the culture is finished, the bacterial concentration of the common triangular flask is measured to be 4.0x10 9 Fu/mL, the fungus concentration of the feeding triangular flask is 8.0X10 9 Fu/mL, the bacterial concentration is about 2.0 times of that of the common triangular flask culture.
After the cultivation is finished, the cultivated seed liquid is inoculated to a fermentation tank respectively in an environment with more mixed bacteria, the seeds cultivated in a common triangular flask are inoculated by a fire ring method, mixed bacteria in the air can be introduced, the seeds cultivated in the improved triangular flask are inoculated by a peristaltic pump in a fed-batch mode, and the condition that the seeds are not infected by bacteria is verified to be fed-batch by the peristaltic pump in a fed-batch mode for many times, so that the effective rate is 100%.
As can be seen from the above examples 2-3, the microbial triangular flask culture apparatus of the present utility model can be used for culturing, and the risk of bacterial contamination during the inoculation process from the triangular flask with less alkali to the fermenter can be effectively increased, and the microbial triangular flask culture apparatus can be used for transferring more fragile strains and inoculating in a worse environment, and the effective rate can reach 100%.
And culturing a plurality of strains such as lactobacillus plantarum, bacillus coagulans, saccharomyces cerevisiae and the like in an environment with more mixed bacteria, and carrying out fed-batch inoculation and fire loop inoculation comparative test, wherein the fire loop inoculation can cause mixed bacteria pollution, and the fed-batch inoculation does not generate mixed bacteria pollution.
The foregoing description is only illustrative of the preferred embodiments of the present utility model and is not to be construed as limiting the scope of the utility model, and it will be appreciated by those skilled in the art that equivalent substitutions and obvious variations may be made using the description and illustrations of the present utility model, and are intended to be included within the scope of the present utility model.
Claims (8)
1. The utility model provides a microorganism triangular flask culture apparatus, includes the triangular flask, its characterized in that, the triangular flask includes glass body (1), glass pipeline (2) and is used for supporting solid glass support (3) of glass pipeline (2), an opening has been seted up on the one side bottle wall of glass body (1), one end of glass pipeline (2) is passed through the opening stretches to the inside bottom of glass body (1), the other end is located the outside of glass body (1), glass pipeline (2) cover is equipped with silica gel transparent hose (7) or running water pipeline (6).
2. The microbial triangular flask culture device according to claim 1, wherein the triangular flask consists of a glass flask body (1) with a conical bottom and an arc-shaped flask mouth (4) above, and a breathable antibacterial sealing film (5) is sleeved on the flask mouth (4).
3. The microbial triangular flask culture device according to claim 1, wherein a sterilizing filter membrane (9) or a sterilizing filter membrane (9) and a syringe (8) or a syringe (8) are arranged on the silica gel transparent hose (7), and the sterilizing filter membrane (9) is detachably connected with the syringe (8).
4. A microbial flask culture apparatus according to claim 3, wherein the pore size of the sterile filter membrane (9) is 0.2-0.25um.
5. The microbial triangular flask culture device according to claim 1, wherein the feeding pipeline (6) comprises a silica gel hose (61) and a stainless steel needle (62), the stainless steel needle (62) is inserted into a feeding interface of a fermentation tank, and the silica gel hose (61) is sleeved on the glass pipeline (2).
6. The microbial triangular flask culture device according to claim 1, wherein the solid glass support (3) is arranged in parallel with the bottom of the glass flask body (1).
7. The microbial triangular flask culture device according to claim 1, wherein one end of the solid glass support (3) is fixedly connected with the glass pipeline (2), and the other end is fixedly connected with the inner bottle wall of the glass bottle body (1).
8. The microbial triangular flask culture device according to claim 1, wherein the glass bottle body (1), the glass pipeline (2) and the solid glass bracket (3) are integrally formed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202223534733.2U CN219652999U (en) | 2022-12-28 | 2022-12-28 | Microorganism triangular flask culture apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202223534733.2U CN219652999U (en) | 2022-12-28 | 2022-12-28 | Microorganism triangular flask culture apparatus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219652999U true CN219652999U (en) | 2023-09-08 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN202223534733.2U Active CN219652999U (en) | 2022-12-28 | 2022-12-28 | Microorganism triangular flask culture apparatus |
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| Country | Link |
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| CN (1) | CN219652999U (en) |
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- 2022-12-28 CN CN202223534733.2U patent/CN219652999U/en active Active
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